A Two Hybrid System Based Yeasy Screen for Agents that Affect DNA Damage Checkpoi

基于两种混合系统的 Yeasy 筛选影响 DNA 损伤检查点的试剂

• 批准号: 7326735
• 负责人: NALIN KUMAR
• 金额: $ 13.54万
• 依托单位: UHV TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7326735
• 关键词: Address; Adjuvant; Affect; Agar; Amplifiers; Antineoplastic Agents; Apoptosis; Automation; Biological Assay; Businesses; Camptothecin; Cancer Control; Cell Cycle Arrest; Cells; Chemotherapy-Oncologic Procedure; Class; Condition; DNA Binding Domain; DNA Damage; Development; Family; Funding; Future; Goals; Growth; Health Sciences; Histidine; Image Analysis; Lead; Libraries; Malignant Neoplasms; Molecular; National Cancer Institute; Normal Cell; Numbers; Pharmaceutical Preparations; Phase; Plasmids; Proteins; Radiation therapy; Reporter; Robotics; Saccharomyces cerevisiae; Sales; Screening procedure; Services; Small Business Technology Transfer Research; Source; System; Techniques; Technology; Testing; Texas; Therapeutic Index; Topoisomerase-I Inhibitor; Toxic effect; United States National Institutes of Health; Universities; Work; Yeasts; base; cancer cell; cancer therapy; chemotherapeutic agent; chemotherapy; cost efficient; drug discovery; high throughput screening; improved; inhibitor/antagonist; innovation; instrument; instrumentation; interest; neoplastic cell; novel; response; small molecule; tool; yeast two hybrid system; Address; Adjuvant; Affect; Agar; Amplifiers; Antineoplastic Agents; Apoptosis; Automation; Biological Assay; Businesses; Camptothecin; Cancer Control; Cell Cycle Arrest; Cells; Chemotherapy-Oncologic Procedure; Class; Condition; DNA Binding Domain; DNA Damage; Development; Family; Funding; Future; Goals; Growth; Health Sciences; Histidine; Image Analysis; Lead; Libraries; Malignant Neoplasms; Molecular; National Cancer Institute; Normal Cell; Numbers; Pharmaceutical Preparations; Phase; Plasmids; Proteins; Radiation therapy; Reporter; Robotics; Saccharomyces cerevisiae; Sales; Screening procedure; Services; Small Business Technology Transfer Research; Source; System; Techniques; Technology; Testing; Texas; Therapeutic Index; Topoisomerase-I Inhibitor; Toxic effect; United States National Institutes of Health; Universities; Work; Yeasts; base; cancer cell; cancer therapy; chemotherapeutic agent; chemotherapy; cost efficient; drug discovery; high throughput screening; improved; inhibitor/antagonist; innovation; instrument; instrumentation; interest; neoplastic cell; novel; response; small molecule; tool; yeast two hybrid system

DESCRIPTION (provided by applicant): Cancer chemotherapy agents frequently introduce DNA damage and, as a consequence, trigger cell cycle arrest or apoptosis, referred to as checkpoint responses. We have developed a two-hybrid-system based yeast assay that detects increased interaction between certain checkpoint proteins following certain DNA-damaging treatments. This phenomenon accompanies checkpoint activation and can be detected by colony growth on selective medium in specially constructed strains of the yeast Saccharomyces cerevisiae. Effective cancer chemotherapy frequently relies on a combination of agents. This application addresses the continued need to identify small molecules that have activity against cancer cells and to characterize novel agents that amplify or modify their effect. We plan to further refine the assay conditions for large-scale robotic screening and, as a proof-of-principle study, to address a sophisticated screening goal: to isolate novel compounds that enhance (or diminish) the checkpoint-activating effect of the established chemotherapy drug camptothecin. It is expected that the usefulness of this agent class of topoisomerase I inhibitors can be enhanced by providing adjuvants that enhance its effect on tumor cells or blunt its toxicity towards normal cells. To this end, the following specific aims will be pursued: 1. To demonstrate the feasibility of developing the assay into a high- throughput screening tool, with special emphasis on screening for agents that modify the camptothecin effect. 2. To modify and test the appropriate instrumentation for demonstration of high throughput capability. 3. To screen a compound library for agents that modify the effects of camptothecin. We plan to screen the National Cancer Institute's Diversity Compound set (1,900 agents) during Phase I that is already available to us. At the successful completion of this project, we will have established and validated a unique, robust, cost efficient and rapid screening system and will have made substantial progress towards isolation of lead compounds that can be developed into amplifiers or adjuvants in camptothecin cancer chemotherapy. Effective cancer chemotherapy frequently relies on a combination of agents. This application addresses the continued need to identify small molecules that have activity against cancer cells and to characterize novel agents that amplify or modify their effect. University of North Texas Health Science Center (UNTHSC) has developed an innovative technique that detects DNA damage caused by chemotherapy agents used in cancer treatment. During this STTR project, UNTHSC will work with a local high technology company (UHV Technologies, Inc.) to further develop this technique for large-scale robotic screening to isolate novel compounds that enhance (or diminish) the effects of the established chemotherapy drug camptothecin. UHV will modify its high throughput cancer drug discovery system developed under previous NIH funding to implement and demonstrate this new technique by screening a compound library for agents that modify the effects of camptothecin.

描述（由申请人提供）：癌症化学疗法经常引入DNA损伤，因此，触发细胞周期停滞或凋亡被称为检查点反应。我们已经开发了一种基于两杂交系统的酵母菌测定法，该测定法可以检测到某些DNA受损处理后某些检查点蛋白之间的相互作用增加。这种现象伴随着检查点的激活，可以通过在酿酒酵母的特殊构建菌株中的选择性培养基上的菌落生长来检测。有效的癌症化学疗法通常依赖于剂的组合。该应用程序解决了持续的需求，以鉴定针对癌细胞活性的小分子，并表征扩大或改变其作用的新型药物。我们计划进一步完善大规模机器人筛选的测定条件，并作为原则研究证明，以解决一个复杂的筛查目标：隔离新型化合物，以增强（或减少）已建立化学疗法药物camptothecin的检查点激活作用。可以通过提供增强其对肿瘤细胞的作用或钝化其对正常细胞的毒性来增强该试剂类药物I抑制剂的有用性。为此，将追求以下特定目标：1。证明将测定法开发为高吞吐量筛选工具的可行性，特别强调筛选改变camptothecin效应的代理。 2。修改和测试适当的仪器以演示高通量能力。 3。筛选一个化合物库的代理，以修改camptothecin的影响。我们计划在第一阶段可以筛选国家癌症研究所的多样性化合物集（1,900个代理商）。成功完成该项目后，我们将建立并验证了一种独特，健壮，具有成本效益和快速筛查系统，并将在隔离铅化合物方面取得了重大进展，这些化合物可以将其发展为camptothecincin癌症化学疗法中的放大器或佐剂。有效的癌症化学疗法通常依赖于剂的组合。该应用程序解决了持续的需求，以鉴定针对癌细胞活性的小分子，并表征扩大或改变其作用的新型药物。北德克萨斯大学健康科学中心（UNTHSC）开发了一种创新技术，该技术检测到癌症治疗中使用的化学疗法剂引起的DNA损伤。在此STTR项目中，UNTHSC将与当地高科技公司（UHV Technologies，Inc。）合作，进一步开发该技术，用于大规模机器人筛选，以隔离新型化合物，从而增强（或减少）已建立的化学疗法药物摄影药的作用。 UHV将修改其在先前的NIH资金下开发的高吞吐量癌症发现系统，以通过筛选一个化合物库的代理，以修改Camptothecin的作用，从而实施和证明这一新技术。